Methods of making and using guidance and navigation control proteins

ABSTRACT

The application provides methods for generating a therapeutic composition. The method includes the steps of providing a cell material comprising a cytotoxic cell, incubating the cell material with a first GNC protein to provide an activated cell composition, wherein the activated cell composition comprises a first therapeutic cell, and formulating the activated cell composition to provide a therapeutic composition, wherein the therapeutic composition is substantially free of exogenous viral and non-viral DNA or RNA. The first GNC protein comprises a first cytotoxic binding moiety and a first cancer targeting moiety, wherein the first cytotoxic binding moiety has a specificity to a first cytotoxic cell receptor and is configured to activate the first cytotoxic cell, and wherein the first cancer targeting moiety has a specificity to a first cancer cell receptor. The first therapeutic cell comprises the first GNC protein bound to the cytotoxic cell through the first cytotoxic cell receptor.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of filing date of U.S. ProvisionalPatent Application No. 62/648,888 filed Mar. 27, 2018, and U.S.Provisional Patent Application No. 62/648,880 filed Mar. 27, 2018, theentire disclosures of which are expressly incorporated by referenceherein.

TECHNICAL FIELD

The present application generally relates to the technical field ofGuidance and Navigation Control (GNC) proteins with multi-specificbinding activities against surface molecules on both immune cells andtumor cells, and more particularly relates to making and using GNCproteins.

BACKGROUND

Cancer cells develop various strategies to evade the immune system. Oneof the underlying mechanisms for the immune escape is the reducedrecognition of cancer cells by the immune system. Defective presentationof cancer specific antigens or lack of thereof results in immunetolerance and cancer progression. In the presence of effective immunerecognition tumors use other mechanisms to avoid elimination by theimmune system. Immunocompetent tumors create suppressivemicroenvironments to downregulate the immune response. Multiple playersare involved in shaping the suppressive tumor microenvironment,including tumor cells, regulatory T cells, Myeloid-Derived Suppressorcells, stromal cells, and other cell types. The suppression of immuneresponse can be executed in a cell contact-dependent format as well asin a contact-independent manner, via secretion of immunosuppressivecytokines or elimination of essential survival factors from the localenvironment. Cell contact-dependent suppression relies on moleculesexpressed on the cell surface, e.g. Programmed Death Ligand 1 (PD-L1),T-lymphocyte-associated protein 4 (CTLA-4) and others (Dunn, Old et al.2004, Adachi and Tamada 2015).

As the mechanisms by which tumors evade recognition by the immune systemcontinue to be better understood, new treatment modalities that targetthese mechanisms have recently emerged. On Mar. 25, 2011, the U. S. Foodand Drug Administration (FDA) approved ipilimumab injection (Yervoy,Bristol-Myers Squibb) for the treatment of unresectable or metastaticmelanoma. Yervoy binds to cytotoxic T-lymphocyte-associated protein 4(CTLA-4) expressed on activated T cells and blocks the interaction ofCTLA-4 with CD80/86 on antigen-presenting cells thereby blocking thenegative or inhibitory signal delivered into the T cell through CTLA-4resulting in re-activation of the antigen-specific T cell leading to, inmany patients, eradication of the tumor. A few years later in 2014 theFDA approved Keytruda (Pembrolizumab, Merck) and Opdivo (Nivolumab,Bristol-Myers Squibb) for treatment of advanced melanoma. Thesemonoclonal antibodies bind to PD-1 which is expressed on activatedand/or exhausted T cells and block the interaction of PD-1 with PD-L1expressed on tumors thereby eliminating the inhibitory signal throughPD-1 into the T cell resulting in re-activation of the antigen-specificT cell leading to again, in many patients, eradication of the tumor.Since then additional clinical trials have been performed comparing thesingle monoclonal antibody Yervoy to the combination of the monoclonalantibodies Yervoy and Opdivo in the treatment of advanced melanoma whichshowed improvement in overall survival and progression-free survival inthe patients treated with the combination of antibodies. (Hodi, Chesneyet al. 2016, Hellmann, Callahan et al. 2018). However, as many clinicaltrials have shown a great benefit of treating cancer patients withmonoclonal antibodies that are specific for one or more immunecheckpoint molecules data has emerged that only those patients with ahigh mutational burden that generates a novel T cell epitope(s) which isrecognized by antigen-specific T cells show a clinical response (Snyder,Makarov et al. 2014). Those patients that have a low tumor mutationalload mostly do not show an objective clinical response (Snyder, Makarovet al. 2014, Hellmann, Callahan et al. 2018).

In recent years other groups have developed an alternate approach thatdoes not require the presence of neoepitope presentation byantigen-presenting cells to activate T cells. One example is thedevelopment of a bi-specific antibody where the binding domain of anantibody which is specific for a tumor associated antigen, e.g., CD19,is linked to an antibody binding domain specific for CD3 on T cells thuscreating a bi-specific T cell engager or BiTe molecule. In 2014, the FDAapproved a bi-specific antibody called Blinatumumab for the treatment ofPrecursor B-Cell Acute Lymphoblastic Leukemia. Blinatumumab links thesingle-chain variable fragment (scFv) specific for CD19 expressed onleukemic cells with the scFv specific for CD3 expressed on T cells(Benjamin and Stein 2016). However, despite an initial response rateof >50% in patients with relapsed or refractory ALL many patients areresistant to Blinatumumab therapy or relapse after successful treatmentwith Blinatumumab. Evidence is emerging that the resistance toBlinatumumab or relapse after Blinatumumab treatment is attributable tothe expression of immune checkpoint inhibitory molecules expressed ontumor cells, such as PD-L1 that drives an inhibitory signal through PD-1expressed on activated T cells (Feucht, Kayser et al. 2016). In a casestudy of a patient who was resistant to therapy with Blinatumumab, asecond round of Blinatumumab therapy was performed but with the additionof a monoclonal antibody, pembrolizumab (Keytruda, Merck). Pembrolizumabspecifically binds to PD-1 and blocks the interaction of Tcell-expressed PD-1 with tumor cell expressed PD-L1, which resulted in adramatic response and reduction of tumor cells in the bone marrow from45% to less than 5% in this one patient (Feucht, Kayser et al. 2016).These results show that combining a bi-specific BiTe molecule with oneor more monoclonal antibodies can significantly increase clinicalactivity compared to either agent alone. Despite the promising outcome,the cost leading to the combined therapy must be high due to multipleclinical trials and the difficulty in recruiting representativepopulations.

Adoptive cell therapy with chimeric antigen receptor T cells (CAR-T) isanother promising immunotherapy for treating cancer. The clinicalsuccess of CAR-T therapy has revealed durable complete remissions andprolonged survival of patients with CD19-positive treatment-refractory Bcell malignancies (Gill and June 2015). However, the cost and complexityassociated with the manufacture of a personalized and geneticallymodified CAR-T immunotherapy has restricted their production and use tospecialized centers for treating relatively small numbers of patients.Cytokine release syndrome (CRS), also known as cytokine storm, isconsidered as the major adverse effect after the infusion of engineeredCAR-T cells (Bonifant, Jackson et al. 2016). In many cases, the onsetand severity of CRS seems to be personally specific to the patient.Current options of mitigating CRS are mainly focused on rapid responseand management care because the option of controlling CRS prior to Tcell infusion is limited.

While the efficacy of CAR-T therapy specific for a CD19-positive B cellmalignancy is now clearly established, the efficacy of CAR-T therapyagainst solid tumors has not been unequivocally demonstrated to date.Currently, many clinical trials are in progress to explore a variety ofsolid tumor-associated antigens (TAA) for CAR-T therapy. Inefficient Tcell trafficking into the tumors, an immunosuppressive tumormicro-environment, suboptimal antigen recognition specificity, and lackof control over treatment-related adverse events are currentlyconsidered as the main obstacles in solid tumor CAR-T therapy (Li, Li etal. 2018). The option of managing the therapeutic effect, as well as anyadverse effect before and after the CAR-T cell infusion, is limited.

SUMMARY

The application provides, among others, methods for generatingtherapeutic compositions containing a guidance and navigation (GNC)proteins, methods for treating cancer conditions using a guidance andnavigation control (GNC) proteins, and therapeutic compositionscontaining GNC proteins or therapeutic cells having cytotoxic cellscoated (or bound) with GNC proteins.

In one aspect, the application provides therapeutic compositions. In oneembodiment, the therapeutic composition comprises a cytotoxic cell, aGNC protein, and a therapeutic cell. The therapeutic cell comprises theGNC protein bound to the cytotoxic cell through the binding interactionwith the cytotoxic cell receptor, and the therapeutic cell compositionis substantially free exogenous of viral and non-viral DNA and RNA.

In one embodiment, the therapeutic composition may further comprise asecond GNC protein, a second therapeutic cell, or a combination thereof,wherein the second therapeutic cell comprises the cytotoxic cells withthe second GNC protein bound thereupon or with both the first and thesecond GNC proteins bound thereupon.

GNC protein includes a cytotoxic binding moiety and a cancer targetingmoiety. The cytotoxic binding moiety has a binding specificity to acytotoxic cell receptor and is configured to activate the cytotoxic cellthrough the binding with the cytotoxic cell receptor. The cancertargeting moiety has a binding specificity to a cancer cell receptor.

In one embodiment, the GNC protein includes a binding domain for T-cellreceptors. Examples T-cell receptor include without limitation CD3,CD28, PDL1, PD1, OX40, 4-1BB, GITR, TIGIT, TIM-3, LAG-3, CTLA4, CD40L,VISTA, ICOS, BTLA, Light, CD30, NKp30, CD28H, CD27, CD226, CD96, CD112R,A2AR, CD160, CD244, CECAM1, CD200R, TNFRSF25 (DR3), or a combinationthereof. In one embodiment, the GNC protein is capable of activating aT-cell by binding the T-cell binding moiety to a T-cell receptor on theT-cell. In one embodiment, the GNC protein is capable of activating aT-cell by binding multiple T-cell binding moieties on the T-cell.

In one embodiment, the GNC protein includes a binding domain for a NKcell receptor. Examples NK cell receptor include, without limitation,receptors for activation of NK cell such as CD16, NKG2D, KIR2DS1,KIR2DS2, KIR2DS4, KIR3DS1, NKG2C, NKG2E, NKG2H; agonist receptors suchas NKp30a, NKp30b, NKp46, NKp80, DNAM-1, CD96, CD160, 4-1BB, GITR, CD27,OX-40, CRTAM; and antagonist receptors such as KIR2DL1, KIR2DL2,KIR2DL3, KIR3DL1, KIR3DL2, KIR3DL3, NKG2A, NKp30c, TIGIT, SIGLEC7,SIGLEC9, LILR, LAIR-1, KLRG1, PD-1, CTLA-4, CD161.

In one embodiment, the GNC protein includes a binding domain for amacrophage receptor. Examples macrophage receptor include, withoutlimitation, agonist receptor on macrophage such as TLR2, TLR4, CD16,CD64, CD40, CD80, CD86, TREM-1, TREM-2, ILT-1, ILT-6a, ILT-7, ILT-8,EMR2, Dectin-1, CD69; antagonist receptors such as CD32b, SIRPa, LAIR-1,VISTA, TIM-3, CD200R, CD300a, CD300f, SIGLEC1, SIGLEC3, SIGLEC5,SIGLEC7, SIGLEC9, ILT-2, ILT-3, ILT-4, ILT-5, LILRB3, LILRB4, DCIR; andother surface receptors such as CSF-1R, LOX-1, CCR2, FRP, CD163, CR3,DC-SIGN, CD206, SR-A, CD36, MARCO.

In one embodiment, the GNC protein includes a binding domain for adendritic cell receptor. Examples dendritic cell receptor include,without limitation, agonist receptors on dendritic cell such as TLR,CD16, CD64, CD40, CD80, CD86, HVEM, CD70; antagonist receptors such asVISTA, TIM-3, LAG-3, BTLA; and other surface receptors such as CSF-1R,LOX-1, CCR7, DC-SIGN, GM-CSF-R, IL-4R, IL-10R, CD36, CD206, DCIR, RIG-1,CLEC9A, CXCR4.

In one embodiment, the GNC protein may include a T-cell binding moietyand a cancer-targeting moiety. In one embodiment, the T-cell bindingmoiety has a binding specificity to a T-cell receptor comprising CD3,CD28, PDL1, PDL2, PD1, OX40, 4-1BB, GITR, TIGIT, TIM-3, LAG-3, CTLA4,CD40L, VISTA, ICOS, BTLA, Light, CD30, CD27, or a combination thereof.In one embodiment, the cancer targeting moiety has a binding specificityto a cancer cell receptor. In one embodiment, the cancer cell receptormay include BCMA, CD19, CD20, CD33, CD123, CD22, CD30, ROR1, CEA, HER2,EGFR, EGFRvIII, LMP1, LMP2A, Mesothelin, PSMA, EpCAM, glypican-3, gpA33,GD2, TROP2, as yet to be discovered tumor associated antigens or acombination thereof.

In one embodiment, the GNC protein may have multi-specific antigenbinding activities to the surface molecules of a T cell and a tumourcell. In one embodiment, the guidance and navigation control (GNC)protein comprises a binding domain for a T cell activating receptor, abinding domain for a tumor associated antigen, a bind domain for animmune checkpoint receptor, and a binding domain for a T cellco-stimulating receptor.

In one embodiment, the binding domain for the tumor associated antigenis not adjacent to the binding domain for the T cell co-stimulatingreceptor. In one embodiment, the binding domain for the T cellactivating receptor is adjacent to the binding domain for the tumorassociated antigen (TAA). The T cell activating receptor may includewithout limitation CD3. The T cell co-stimulating receptor may includewithout limitation 4-1BB, CD28, OX40, GITR, CD40L, ICOS, Light, CD27,CD30, or a combination thereof. The immune checkpoint receptor mayinclude without limitation PD-L1, PD-1, TIGIT, TIM-3, LAG-3, CTLA4,BTLA, VISTA, PDL2, or a combination thereof.

The tumor associated antigen (TAA) may include without limitation ROR1,CD19, EGFRVIII, BCMA, CD20, CD33, CD123, CD22, CD30, CEA, HER2, EGFR,LMP1, LMP2A, Mesothelin, PSMA, EpCAM, glypican-3, gpA33, GD2, TROP2, ora combination thereof. In one embodiment, the tumor associated antigenmay be ROR1. In one embodiment, the tumor associated antigen may beCD19. In one embodiment, the tumor associated antigen may be EGFRVIII.

In one embodiment, the guidance and navigation control (GNC) protein maybe an antibody or an antibody monomer or a fragment thereof. In oneembodiment, the GNC protein may be a tri-specific antibody. In oneembodiment, the GNC protein may be a tetra-specific antibody. In oneembodiment, the GNC protein includes Fc domain or a fragment thereof.Any Fc domain from an antibody may be used. Example Fc domains mayinclude Fc domains from IgG, IgA, IgD, IgM, IgE, or a fragment or acombination thereof. Fc domain may be natural or engineered. In oneembodiment, the Fc domain may contain an antigen binding site.

In one embodiment, the GNC protein comprises a bi-specific antibody, atri-specific antibody, a tetra-specific antibody, or a combinationthereof yielding up to eight binding motifs on the GNC protein. Examplesof antibodies, antibody monomers, antigen-binding fragment thereof aredisclosed herein. In one embodiment, GNC proteins may include animmunoglobulin G (IgG) moiety with two heavy chains and two lightchains, and at least two scFv moieties being covalently connected toeither C or N terminals of the heavy or light chains. The IgG moiety mayprovide stability to the scFv moiety, and a tri-specific GNC protein mayhave two moieties for binding the surface molecules on T cells.

In one embodiment, the guidance and navigation control (GNC) protein maybe an antibody. In one embodiment, the tumor associated antigencomprises ROR1, CD19, or EGRFVIII. In on embodiment, the T cellactivating receptor comprises CD3 and the binding domain for CD3 may belinked to the binding domain for the tumor associated (TAA) antigenthrough a linker to form a CD3-TAA pair. In one embodiment, the IgG Fcdomain may intermediate the CD3-TAA pair and the binding domain for theimmune checkpoint receptor. In one embodiment, the immune checkpointreceptor may be PD-L1.

The linker may be a covalent bond or a peptide linker. In oneembodiment, the peptide linker may have from about 2 to about 100 aminoacid residues.

In on embodiment, the guidance and navigation control (GNC) protein hasa N-terminal and a C-terminal, comprising in tandem from the N-terminalto the C-terminal, the binding domain for CD3, the binding domain forEGFRVI, IgG Fc domain, the bind domain for PD-L1, and the binding domainfor 41-BB. In one embodiment, the guidance and navigation control (GNC)protein has a N-terminal and a C-terminal, comprising in tandem from theN-terminal to the C-terminal, the binding domain for 4-1BB, the bindingdomain for PD-L1, IgG Fc domain, the bind domain for ROR1, and thebinding domain for CD3. In one embodiment, the guidance and navigationcontrol (GNC) protein has a N-terminal and a C-terminal, comprising intandem from the N-terminal to the C-terminal, the binding domain forCD3, the binding domain for CD19, IgG Fc domain, the bind domain forPD-L1, and the binding domain for 4-1BB.

In one embodiment, the GNC protein comprises an amino acid having apercentage homology to SEQ ID NO. 50, 52, 80, 82, 84, 86, 88, 90, 92,94, 96, 98, 100, 102, 104, 106, 108, and 110. The percentage homology isnot less than 70%. 80%, 90%, 95%, 98% or 99%.

In another aspect, the application provides nucleic acid sequencesencoding the GNC protein or its fragments disclosed thereof. In oneembodiment, the nucleic acid has a percentage homology to SEQ ID NO. 49,51, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, and109. The percentage homology is not less than 70%. 80%, 90%, 95%, 98% or99%.

In another aspect, the application provides methods for generating atherapeutic composition. In one embodiment, the method may include thesteps of providing a cell material comprising a cytotoxic cell,incubating the cell material with a first GNC protein to provide anactivated cell composition, and formulating the activated cellcomposition to provide a therapeutic composition. The activated cellcomposition contains a first therapeutic cell. The first therapeuticcell comprises the first GNC protein bound to the cytotoxic cell throughthe binding interaction with the first cytotoxic cell receptor. Thetherapeutic composition is substantially free of exogenous viral andnon-viral DNA or RNA.

In one embodiment, the cell material may include or be derived fromPBMC.

The first GNC protein may include a first cytotoxic binding moiety and afirst cancer targeting moiety. The first cytotoxic binding moiety has aspecificity to a first cytotoxic cell receptor and is configured toactivate the first cytotoxic cell through the binding with the firstcytotoxic cell receptor. The first cancer targeting moiety has aspecificity to a first cancer cell receptor.

In one embodiment, the method may repeat the incubating step byincubating a second GNC protein with the activated cell composition. Thesecond GNC protein comprising a second cytotoxic binding moiety and asecond cancer targeting moiety, the second cytotoxic binding moiety hasa specificity to a second cytotoxic cell receptor, and the second cancertargeting moiety has a specificity to a second cancer cell receptor. Theactivated cell composition comprises a second therapeutic cell, and thesecond therapeutic cell comprises the second GNC protein bound to thecytotoxic cell or the first therapeutic cell through the bindinginteraction with the second cytotoxic cell receptor.

In one embodiment, the first and the second cancer-targeting moietyindependently has a specificity for CD19, PDL1, or a combinationthereof. In one embodiment, the first and the second cytotoxic bindingmoiety independently has a specificity for CD3, PDL1, 41BB, or acombination thereof.

The method may further include the repeated incubating steps byincubating additional GNC proteins with the activated composition. Theadditional GNC proteins may be a third GNC protein, a fourth GNCprotein, etc. to provide addition therapeutic cells, each having theadditional protein bound to the cytotoxic cell.

The first, second, and the additional GNC protein may be the same or maybe different. The therapeutic cells may have one GNC protein, multiplesame GNC proteins, or multiple different GNC proteins bound thereupon.In one embodiment, the therapeutic cell may have the first GNC proteinbound thereupon. In one embodiment, the therapeutic cell may have boththe first and the second GNC proteins bound thereupon. In oneembodiment, the therapeutic cell may have the first, the second and theadditional GNC proteins bound thereupon.

In one embodiment, the therapeutic cell comprises the cytotoxic cellhaving at least one bound GNC protein. In one embodiment, thetherapeutic cell comprises the cytotoxic cell having at least 10, 20,50, 100, 200, 300, 400 bound GNC proteins.

The therapeutic composition may include the first therapeutic cell, thefirst GNC protein, the cytotoxic cell, or a combination thereof. In oneembodiment, the therapeutic composition may include the secondtherapeutic cell, the second GNC protein, comprises the firsttherapeutic cell, the first GNC protein, the cytotoxic cell, or acombination thereof. In one embodiment, the therapeutic composition mayinclude additional GNC proteins and additional therapeutic cells.

In one embodiment, the incubating step may serve to expand thetherapeutic cells. In one embodiment, expanding the therapeutic cell mayinclude incubating the therapeutic cells with an additional amount ofthe GNC protein to provide an expanded cell population. In oneembodiment, the expanded cell population comprises at least 10², atleast 103, at least 10⁴, at least 10⁵, at least 10⁶, at least 10⁷, atleast 10⁸, at least 10⁹, at least 10¹⁰ cells per ml. In one embodiment,the expanded cell population comprises the GNC bound cell, the GNCprotein, the cytotoxic cell, or a combination thereof. In oneembodiment, in order to deplete PD-1+ T cells, a GNC protein may beadded to the expansion culture that redirects killing to PD-1+ T cellstherefore resulting in reduction in PD-1+ exhausted T cells. In oneembodiment, in order to preferentially support PD-1+ T cells, a GNCprotein may be added to the expansion culture that relieves checkpointsignaling through PD-1 on T cells therefore resulting in functionalimprovement of PD-1+ T cells. In one embodiment, in order to isolate4-1BB mediated co-stimulation through 3^(rd) gen CAR-T, a GNC proteinmay be added to the expansion culture that redirects killing to 4-1BB+ Tcells or resulting in therapeutic composition with controlling level of4-1BB stimulation in the therapeutic cells, such as CAR-T cells.

In one embodiment, the cancer targeting moiety has the specificityagainst B cell, and the therapeutic composition is substantially free ofB cell. Therefore, the methods disclosed herein couple the activationand purification functions for the therapeutic cells, which allows themethods to produce B cell free therapeutic composition without the needto introduce any foreign materials (such as beads) nor any foreigngenetic materials (such as viral and non-viral DNA or RNA vectors).

In one embodiment, the ratio of the GNC protein and the cytotoxic cellis at least 30 to 1 when incubating the cell material with the GNCprotein.

In one embodiment, the therapeutic composition may include at least 10⁷cells per ml.

In a further aspect, the application provides methods for using guidanceand navigation control (GNC) proteins for cancer treatment. In oneembodiment, the method of treating a subject having a cancer, comprisesproviding a cytotoxic cell, combining a GNC protein with the cytotoxiccell to provide a therapeutic cell, optionally expanding the therapeuticcell to provide an expanded cell population, and administering thetherapeutic cell or the expanded cell population to the subject.

In one embodiment, the method include the step of providing a cellmaterial comprising a cytotoxic cell, incubating the cell material witha first GNC protein to provide an activated cell composition, whereinthe activated cell composition comprises a first therapeutic cell,formulating the activated cell composition to provide a therapeuticcomposition, wherein the therapeutic composition is substantially freeexogenous of viral and non-viral DNA or RNA, and administering thetherapeutic composition to the subject.

In one embodiment, the method may further include the steps ofincubating a second GNC protein with the activated cell composition toprovide the activated cell composition further comprising a secondtherapeutic cell. In one embodiment, the method may further include thestep of incubating additional GNC proteins with the activated cellcomposition to provide the activated cell composition further comprisingadditional therapeutic cells.

In one embodiment, the method may further comprise isolating thecytotoxic cell from peripheral blood mononuclear cells (PBMC) beforeproviding the cytotoxic cell. In one embodiment, the method may furthercomprise isolating the peripheral blood mononuclear cells (PBMC) from ablood. In one embodiment, the blood is from the subject. In oneembodiment, the blood is not from the subject. In one embodiment, thecytotoxic cells may be from the patient that is under treatment or adifferent individual, such as a universal donor.

In one embodiment, the cytotoxic cell may be an autologous T cell, analloreactive T cell, or a universal donor T cell. In one embodiment,when autologous donor T cells are used, in order to prevent infusion ofcontaminating cancer cells, a GNC protein may be added to the expansionculture that redirects killing to tumor antigens, example tumor antigenmay include CD19 for B cell malignancies, Epcam for Breast carcinoma,MCP1 for melanoma.

In one embodiment, the method includes steps of providing a blood fromthe subject, isolating peripheral blood mononuclear cells (PBMC) fromthe blood, isolating a cytotoxic cell from the PBMC, combining a GNCprotein with the cytotoxic cell to provide a therapeutic cell,optionally expanding the therapeutic cell to provide an expanded cellpopulation, and administering the therapeutic cell or the expanded cellpopulation to the subject.

In one embodiment, the method further comprises administering additionalGNC protein to the subject after administering the therapeuticcomposition to the subject. In one embodiment, the cytotoxic cell mayinclude CD3+ T cell, NK cell, or a combination thereof.

In one embodiment, the isolating of the cytotoxic cell comprisesisolating at least one subpopulation of cytotoxic cells to provide thetherapeutic T cells. In one embodiment, the subpopulation of cytotoxiccells comprises CD4+ cells, CD8+ cells, CD56+ cells, CD69+ cells,CD107a+ cells, CD45RA+ cells, CD45RO+ cells, CD2+ cells, CD178+ cells,Granzyme+ cells, or a combination thereof.

In one embodiment, the combining of a GNC protein with the cytotoxiccell comprises incubating the GNC protein with the cytotoxic cell for aperiod of time from about 2 hours to about 14 days, from about 1 day toabout 7 days, from about 8 hours to about 24 hours, from about 4 days toabout 7 days, or from about 10 days to about 14 days. In one embodiment,the incubating period may be more than 14 days. In one embodiment, theincubating period may be less than 2 hours.

In one embodiment, the ratio between the GNC protein and the cytotoxiccell is at least 600 to 1, 500 to 1, 400 to 1, 300 to 1, 200 to 1, 100to 1, or 1 to 1. In one embodiment, the ratio between the GNC proteinand the cytotoxic cell is from about 1 to 1, 10 to 1, 100 to 1, or toabout 1000 to 1 ratio.

In one embodiment, the method may further comprise evaluatingtherapeutic efficacy after the administering step. In one embodiment,the evaluating therapeutic efficacy includes checking one or morebiomarkers of the cancer, monitoring the life span of the therapeuticcells, or a combination thereof. In one embodiment, evaluatingtherapeutic efficacy comprises checking one or more biomarkers of thecancer, monitoring the life span of the therapeutic cells, or acombination thereof. In one embodiment, the biomarker comprises a tumorantigen, release of cytokines e.g., gamma interferon, IL-2, IL-8, and/orchemokines, and/or CD markers on the surface of various cell types e.g.,CD69, PD-1, TIGIT, and/or mutated nucleic acid released into thebloodstream by tumors upon death, circulating tumor cells and theirassociated nucleic acid, or exosome associated nucleic acid, hostinflammatory mediators, or tumor derived analytes, or a combinationthereof. In one embodiment, the biomarker comprises a tumor antigen,tumor-associated apoptotic bodies, small molecule metabolites, releaseof cytokines, lymphocyte surface marker expression,phosphorylated/dephosphorylated signaling molecules, transcriptionfactors, or a combination thereof.

The method disclosed herein is free of the step of transfecting thecytotoxic cell with a DNA vector or a viral vector. In one embodiment,the therapeutic cell or the expanded cell population is substantiallyfree of a DNA vector or a viral vector.

The method may be used to treat a human subject suffering from cancer.In one embodiment, the cancer comprises cells expressing ROR1, CEA,HER2, EGFR, EGFRvIII, LMP1, LMP2A, Mesothelin, PSMA, EpCAM, glypican-3,gpA33, GD2, TROP2, BCMA, CD20, CD33, CD123, CD22, CD30, CD19, as yet tobe identified tumor associated antigens, or a combination thereof. Inone embodiment, the method may be used to treat mammals.

Varieties of cancer may be treated using the methods disclosed herein.Example cancers includes without limitation breast cancer, colorectalcancer, anal cancer, pancreatic cancer, gallbladder cancer, bile ductcancer, head and neck cancer, nasopharyngeal cancer, skin cancer,melanoma, ovarian cancer, prostate cancer, urethral cancer, lung cancer,non-small lung cell cancer, small cell lung cancer, brain tumor, glioma,neuroblastoma, esophageal cancer, gastric cancer, liver cancer, kidneycancer, bladder cancer, cervical cancer, endometrial cancer, thyroidcancer, eye cancer, sarcoma, bone cancer, leukemia, myeloma or lymphoma.

In one embodiment, the method may further include administering aneffective amount of a therapeutic agent after the administering thetherapeutic cell or the expanded cell population to the subject. In oneembodiment, the therapeutic agent comprises a monoclonal antibody, achemotherapy agent, an enzyme, a protein, a co-stimulator, or acombination thereof. In one embodiment, the co-stimulator is configuredto increase the amount of cytotoxic T cells in the subject.

The application further provides a solution comprising an effectiveconcentration of the GNC protein. In one embodiment, the solution isblood plasma in the subject under treatment. In one embodiment, thesolution includes the GNC protein bound cells. In one embodiment, thesolution includes a GNC cluster including a GNC protein, a T-cell boundto the T-cell binding moiety of the GNC protein, and a cancer cell isbound to the caner-targeting moiety of the GNC protein.

The objectives and advantages of the present application will becomeapparent from the following detailed description of preferredembodiments thereof in connection with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other features of this disclosure will become morefully apparent from the following description and appended claims, takenin conjunction with the accompanying drawings.

Understanding that these drawings depict only several embodimentsarranged in accordance with the disclosure and are, therefore, not to beconsidered limiting of its scope, the disclosure will be described withadditional specificity and detail through use of the accompanyingdrawings, in which:

FIG. 1 shows a GNC protein comprising four antigen-specific bindingdomains in an antibody structure with targeting specificity to CD19positive cells;

FIG. 2 illustrates that a tetra-specific GNC antibody mediatesmulti-specific binding between a T cell and a tumor cell;

FIG. 3 is a flowchart comparing manufacturing processes for GNC-T celltherapy (left) and CAR-T cell therapy (right);

FIG. 4 is a diagram showing sources of cell material for preparingGNC-activated therapeutic cell composition;

FIG. 5 is a diagram showing sources of selected T cells for preparingGNC-activated therapeutic composition;

FIG. 6 is a diagram showing the preparation of GNC-activated therapeuticT cell composition;

FIG. 7 is a diagram showing the incubating and formulating steps forpreparing the first GNC-activated T cells for GNC-T cell therapy;

FIG. 8 shows that GNC proteins (SI-35E class) induce IL-2 secretion fromPBMC;

FIG. 9 shows that GNC proteins (SI-35E class) induce granzyme Bsecretion from PBMC;

FIG. 10 shows that GNC proteins (SI-35E class) induce expression of theactivation marker CD69 on CD4+ T cells;

FIG. 11 shows that GNC proteins (SI-35E class) induce expression of theactivation marker CD69 on CD8+ T cells;

FIG. 12 shows that GNC proteins (SI-35E class) induce expression of theactivation marker CD69 on CD56+NK cells;

FIG. 13 shows that GNC proteins (SI-35E class) induce expression of themarker of cytotoxic degranulation CD107a on CD4+ T cells;

FIG. 14 shows that GNC proteins (SI-35E class) induce expression of themarker of cytotoxic degranulation CD107a on CD8+ T cells;

FIG. 15 shows that GNC proteins (SI-35E class) induce expression of themarker of cytotoxic degranulation CD107a on CD56+NK cells;

FIG. 16 shows that GNC proteins (SI-35E class) activate CD3+ T cells toproliferate;

FIG. 17 shows that GNC proteins (SI-35E class) activate CD3+ T cells tosecrete gamma interferon;

FIG. 18 shows that GNC proteins (SI-35E class) activate naïveCD8+/CD45RA+ T cells to proliferate;

FIG. 19 shows that GNC proteins (SI-35E class) activate naïveCD8+/CD45RA+ T cells to secrete gamma interferon;

FIG. 20 shows Images of GNC activated cell growth in 6-well G-Rex platesover time;

FIG. 21 shows the example process of making the therapeutic compositionas disclosed thereof (A), and cell viability of PBMC, GET, and GNC-Tcells after thawing (B);

FIG. 22 shows the result of flow cytometry analyses of PBMC-derived, thefirst GNC (SI-38E17)-activated therapeutic cell composition (Product A)(22A), the second GNC (SI-38E17)-coated therapeutic cell composition(Product B) (22B), and input PBMC cell material (22C).

FIG. 23 shows GNC-T therapeutic cell composition of GET cells andformulated GNC-T cells from G-Rex 100M bioreactor after thawing;

FIG. 24 shows the result of RTCC of CHO-ROR1 cells by using GNC (SI-35Eclass)-coated PBMC cells;

FIG. 25 shows kinetics of PBMC-derived, SI-38E17 GNC-activatedtherapeutic cells on killing precursor B cell leukemia Kasumi over time;

FIG. 26 shows efficacy of killing Nalm-6, MEC-1, Daudi, and Jurkat cellsby using PMBC-derived, SI-38E17 GNC-activated therapeutic cells; and

FIG. 27 shows the killing of Nalm-6, MEC-1, Daudi, and Jurkat leukemiccells by using PBMC-derived, SI-38E17 GNC-activated therapeutic cells ina spike-in model.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented herein. It will be readily understood that the aspects of thepresent disclosure, as generally described herein, and illustrated inthe Figures, can be arranged, substituted, combined, separated, anddesigned in a wide variety of different configurations, all of which areexplicitly contemplated herein.

In one embodiment, the guidance navigation control (GNC) proteins arecharacterized by their composition of multiple antigen-specific bindingdomains (AgBDs) and by their ability of directing T cells (or othereffector cells) to cancer cells (or other target cells such as bystandersuppressor cells) through the binding of multiple surface molecules on aT cell and a tumor cell. In one embodiment, GNC proteins are composed ofMoiety 1 for binding at least one surface molecule on a T cell andMoiety 2 for binding at least one surface antigen on a cancer cell asshown in TABLE 1. FIG. 1 shows the structure of an exampletetra-specific GNC antibody comprising AgBDs for binding to both a Tcell expressing CD3, PD-L1, and/or 4-1BB and a target B cell expressingCD19, as illustrated in FIG. 2.

In a T cell therapy, the cytotoxic T cells are regulated by T cellreceptor complex proteins, as well as co-stimulation signaling proteinsvia either agonist receptors or antagonist receptors on their surface.To regulate this signaling, as well as the interaction between a T celland a cancer cell, multiple AgBDs may compose Moiety 1 and Moiety 2,respectively. Examples of molecules that can be targeted by agonistic orantagonistic binding domains in Moiety 1 and 2 are shown in TABLE 1. Inone embodiment, the GNC proteins may have at least one linker to linkMoiety 1 and Moiety 2. In one example GNC protein, any linker moleculecan be used to link two or more AgBDs together either in vitro or invivo by using complementary linkers of DNA/RNA or protein-proteininteractions, including but not limited to, that of biotin-avidin,leucine-zipper, and any two-hybrid positive protein. In someembodiments, the linkers may be an antibody backbone structure orantibody fragments, so that GNC protein and GNC antibody may have thesame meaning, e.g. the structure of the example tetra-specific GNCantibody in FIG. 1.

GNC proteins or antibodies are capable of directing a T cell to a cancercell, in vivo or ex vivo, through the binding function of multiple AgBDs(FIG. 2). The T cells may be derived from the same patient or differentindividuals, and the cancer cell may exist in vivo, in vitro, or exvivo. The examples provided in the present application enable GNCproteins as a prime agent in a T cell therapy, i.e. GNC-T cell therapy,for activating and controlling cytotoxic T cells ex vivo, prior toadoptive transfer.

The present application relates to methods of making GNC-activatedtherapeutic cell composition. Multiple AgBDs can be divided into Moiety1 and Moiety 2 due to their interface with a T cell and a cancer cell,respectively (TABLE 1). A GNC protein with two AgBDs may simultaneouslybind to a surface molecule, such as CD3 on a T cell, and a tumorantigen, such as ROR1 on a tumor cell, for re-directing the T cell tothe tumor cell.

The addition of a third AgBD, for example, one that specifically bindsto 41BB, may help enhance anti-CD3-induced T cell activation because41BB is a co-stimulation factor and the binding stimulates its agonistactivity to activated T cells. The addition of a fourth AgBD to a GNCprotein, for example, one that specifically binds to PD-L1 on a tumorcell, may block the inhibitory pathway of PD-L1 on tumor cells or thatis mediated through its binding to PD-1 on the T cells.

In some embodiments, with these basic principles, GNC proteins areconstructed to acquire multiple AgBDs specifically for binding unequalnumbers of T cell antagonists and agonists, not only to re-directactivated T cells to tumor cells but also to control their activity invivo (TABLE 2). Therefore, in some embodiments, GNC proteins may bebi-specific, tri-specific, tetra-specific, penta-specific,hexa-specific, hepta-specific, or octa-specific proteins.

In one embodiment, the application relates to a GNC-T cell therapy whereGNC proteins are used to expand the T cells ex vivo prior to adoptivetransfer (FIG. 3). The ex vivo priming of autonomous T cells providesthe cytotoxic T cells guidance and navigation control. For example,peripheral blood mononuclear cells (PBMC) or specific types of cellpopulations within PBMC e.g., CD8+, CD45RO+ memory T cells may beisolated and primed ex vivo by GNC proteins. These expanded cytotoxic Tcells can be formulated and infused back to the patient through adoptivetransfer. While attacking the cancer in vivo, additional GNC proteinsmay be infused into the patient for managing the efficacy and lifespanof cytotoxicity. Thus, GNC-T cell therapy is different from GNCprotein-based immunotherapy, where GNC proteins are directlyadministered into patients. However, GNC-T cell therapy does not ruleout the direct administration of GNC proteins for managing the efficacyof infused cytotoxic T cells in vivo in a controlled manner. AdditionalGNC protein can both promote cytolytic activity and encourage T cellproliferation dependent of the configuration of AgBDs.

In one aspect, the application relates to the production of therapeuticGNC-T cells. In comparison with and to distinguish from the productionof therapeutic CAR-T cells, their general processes are shown in FIG. 3,for comparison purpose. In CAR-T therapy, cell material, for examplepatient leukocytes, are collected by apheresis, and a subset of CD3+ Tcells is selected and activated to facilitate gene transfer to thecellular material, which is then expanded in number by the introductionof foreign material scaffold for support to the T cell populations, forexample, by using anti-CD3/anti-CD28 antibody coated beads.Advantageously, GNC-T cell material does not require the introduction ofscaffold impurities for T cell expansion from patient leukocytes.

The CAR-T therapy cellular material must undergo the gene transfer thatinvolves the preparation and transfection of CAR-T vector DNA, whichresults in genetically modifying the genome of the T cells. Furthermore,these genetically modified T cells may undergo another round of T cellexpansion before being transferred back into the patient. The randomintegration of CAR-T vector DNA carries a risk of transformation of theT cells leading to primary leukemogenesis or introduction of the CAR-Tvector to leukemia cells increasing the risk of relapse by mechanism ofinternal sequestration of the CAR target antigen (Zhang, Liu et al.2017).

In contrast, GNC-T cell therapy has the advantages of not involving thetransfection of any vector DNA, therefore there is no risk of geneticmodification prior to adoptive transfer, which provides one of thesignificant advantages and technical improvements over the existingCAR-T therapy. Besides the advantage of GNC-T cell therapy being free ofexogenous generic material contamination and cancer risk, the efficacyof GNC-T cell therapy may be improved when PBMC or different T cellsubsets are being primed and activated ex vivo as shown in FIGS. 5 & 6.Similar approaches have been explored in the use of CAR-T therapy, whereselected specific ratios of some subsets of T cells may be transferredback to the patient (Turtle, Hanafi et al. 2016, Turtle, Hanafi et al.2016).

In some embodiments, it may be beneficial to remove leukemia or othercancer cells from the cellular material prior to cell expansion (FIG.7). The PBMC of a patient with circulating leukemic cells, in particularfrom B cell malignancy, may profoundly alter the cellular compositionand thus affect the suitability of the final therapeutic cellularproducts. For example, a high level of circulating leukemic blast cells(greater that 10% of WBC) may require a depletion of leukemic cellsprior to GNC mediated cell expansion. The percentage of leukemic cellsin the PBMC derived from a patient may be reduced by using cellfractionation methods. These methods may include steps involving densitygradient separation, or immunofluorescent cell separation or fluorescentactivated cells sorting, immunomagnetic cell separation, or microfluidicflow chambers methods. These methods may be preceded by or followcentrifugation, cell washing, incubation, or temperature modulation.These methods may utilize non-cellular substrates (magnetic beads,Plastic, polymers), modification of non-cellular substrates (protein,antibodies, charge state), antibody treatment, multiple antibodytreatments, multi-specific antigen binding proteins and cell surfaceantigen-based cell coupling. These methods may use enzymatic digestionor, ionic chelation, or mechanical agitation or cell vessel rotation.The method for reduction of leukemic blasts may utilize antibody drugconjugates, or leukemia sensitizing agents. The method may consist of acombination of these approaches.

In one embodiment, to enable the production of therapeutic T cellsprimed (or coated or bound) with GNC proteins, a tetra-specific antibodyis produced and used as the GNC protein. In one embodiment, thetetra-specific antibody/GNC protein comprises 4 different bindingdomains linked by antibody fragments as its backbone. One binding domainis specific for CD3 on T cells, a second binding domain is specific fora tumor associated antigen, including but not limited to ROR1, CEA,HER2, EGFR, EGFRvIII, LMP1, LMP2A, Mesothelin, PSMA, EpCAM, glypican-3,gpA33, GD2, TROP2, BCMA, CD19, CD20, CD33, CD123, CD22, CD30, and athird and fourth binding domains are specific for two distinct immunecheckpoint modulators such as PD-L1, PD-L2, PD-1, OX40, 4-1BB, GITR,TIGIT, TIM-3, LAG-3, CTLA4, CD40L, VISTA, ICOS, BTLA, Light, etc.

Without being bound by theory, the advantages of GNC protein-mediatedGNC-T cell therapy over conventional CAR-T therapies include, but arenot limited to, first, that inclusion of an IgG Fc domain may confer thecharacteristic of a longer half-life in serum compared to a bi-specificBiTe molecule; second, that inclusion of two binding domains specificfor immune checkpoint modulators may inhibit the suppressive pathwaysand engage the co-stimulatory pathways at the same time; third, thatcross-linking CD3 on T cells with tumor associated antigens re-directsand guides T cells to kill the tumor cells without the need of removingT cells from the patient and genetically modifying them to be specificfor the tumor cells before re-introducing them back into the patient,also known as chimeric antigen receptor T cells (CAR-T) therapy; andfourth, that GNC protein-mediated antibody therapy or T cell therapydoes not involve genetic modification of T cells, the latter of whichmay carry the risk of transforming modified T cells to clonal expansion,i.e. T cell leukemia.

The present disclosure may be understood more readily by reference tothe following detailed description of specific embodiments and examplesincluded herein. Although the present disclosure has been described withreference to specific details of certain embodiments thereof, it is notintended that such details should be regarded as limitations upon thescope of the disclosure.

EXAMPLES

While the following examples are provided by way of illustration onlyand not by way of limitation. Those of skill in the art will readilyrecognize a variety of non-critical parameters that could be changed ormodified to yield essentially the same or similar results.

Example 1. GNC Proteins and Tetra-Specific GNC Antibodies

In the present application, the examples of GNC proteins are classes oftetra-specific GNC antibodies, of which 4 AgBDs are covalently linkedusing an IgG antibody as its backbone (FIG. 1). From the N-terminal ofthis protein, the first scFv is linked to the Fab domain of the constantdomains C_(H)1, 2, and 3 of IgG antibody which is then linked to anotherscFv at the C-terminal. Because each of the scFv domains displayindependent binding specificity, linking of these AgBDs does not need tobe done using the constant domains of an IgG antibody. Structured as atetra-specific GNC antibody, a GNC protein can directly bind totumor-associated antigen (TAA) and engage the host endogenous T cells tokill tumor cells independent of tumor antigen presentation by MHC to theantigen specific T cell receptors (FIG. 2). As shown in FIG. 1, CD19 isa TAA targeting CD19 positive B cells and tumor cells. In addition,PD-L1 is an example of the immune checkpoint modulating component fortetra-specific GNC antibodies that may overcome the immunosuppressivetumor microenvironment and fully activate the exhausted T cells withinthe tumor microenvironment.

Of tetra-specific GNC antibodies, the SI-35E class comprises targets ananti-human CD3 binding domain (SEQ IDs 1-4), an anti-human PD-L1 (SEQIDs 5-12), an anti-human 4-1BB (SEQ IDs 13-24), and targets a human ROR1(SEQ IDs 25-32), i.e. a TAA. In this context, the classes of SI-38E andSI-39E target CD19 (SEQ IDs 47-50) and EGFR (SEQ ID 51-54),respectively.

To construct tetra-specific GNC antibodies, AgBDs were converted to scFvand VLVH for placement at the N-terminal Domain 1 (D1) or scFv and VHVLfor placement at the C-terminal Domains 3 (D3) and 4 (D4) of the GNCprotein. All scFv molecules described herein contain a 20 amino acidflexible gly-gly-gly-gly-ser (G4S) X4 linker that operably links the VHand VL, regardless of the V-region orientation (LH or HL). The remainingposition in the tetra-specific GNC antibody, Domain 2 (D2), consists ofan IgG1 heavy chain, VH-CH1-Hinge-CH2-CH3, and its corresponding lightchain, VL-CL, which can be either a kappa or lambda chain. D1 and D2 aregenetically linked through a 10 amino acid (G4S)×2 linkers, as are D2,D3 and D4 resulting in a contiguous ˜150 kDa heavy chain monomerpeptide. When co-transfected with the appropriate light chain, the finalsymmetric tetra-specific GNC peptide can be purified through the IgG1 Fc(Protein A/Protein G) and assayed to assess functional activity. Heavyand light chain gene “cassettes” were previously constructed such thatV-regions could be easily cloned using either restriction enzyme sites(HindIII/NhelI for the heavy chain and HindIII/BsiWI for the lightchain) or “restriction-free cloning” such as Gibson Assembly (SGI-DNA,La Jolla, Calif.), Infusion (Takara Bio USA) or NEBuilder (NEB, Ipswich,Mass.), the latter of which was used here.

The tetra-specific GNC antibodies can be produced through a process thatinvolves design of the intact molecule, synthesis and cloning of thenucleotide sequences for each domain, expression in mammalian cells andpurification of the final product. Herein, nucleotide sequences wereassembled using the Geneious 10.2.3 software package (Biomatters,Auckland, NZ) and broken up into their component domains for genesynthesis (Genewiz, South Plainsfield, N.J.). In this example, SI-35E18(SEQ ID 65 and 67) was split into its component domains where theanti-41BB scFv, VL-VH, occupies D1, anti-human PD-L1 clone PL230C6occupies D2 (Fab position), anti-human ROR1 Ig domain-specific clone323H7 VHVL scFv occupies D3, and anti-human CD3 scFv, VHVL, occupies theC-terminal D4. Using NEBuilder web-based tools, 5′ and 3′ nucleotideswere appended to each of the domains depending on their position in thelarger protein so that each domain overlaps its flanking domains by20-30 nucleotides which direct site-specific recombination, thusgenetically fusing each domain in a single gene assembly step. Due tothe high number of homologous regions in the tetra-specific nucleotidesequence, the N-terminal domains 1 and 2 are assembled separately fromthe C-terminal D3 and D4. The N- and C-terminal fragments were thenassembled together in a second NEBuilder reaction. A small aliquot wastransformed into E. coli DH10b (Invitrogen, Carlsbad, Calif.) and platedon TB+carbenicillin 100 ug/ml plates (Teknova, Hollister, Calif.) andincubated at 37° C. overnight. Resultant colonies were selected and 2 mLovernight cultures inoculated in TB+carbenicillin. DNA was prepared(Thermo-Fisher, Carlsbad, Calif.) from overnight cultures andsubsequently sequenced (Genewiz, South Plainsfield, N.J.) usingsequencing primers (Sigma, St. Louis, Mo.) flanking each domain. All DNAsequences were assembled and analyzed in Geneious.

In another tetra-specific GNC protein, SI-38E17 targeting human CD19(SEQ IDs 47-50), multiple AgBDs carry an anti-human 4-1BB (scFv 466F6,SEQ IDs 17-20) as well as an anti-human PD-L1 (scFv PL221G5 SEQ IDs9-13), and an anti-human CD3 binding domain (SEQ IDs 1-4). The methodsand procedures for producing this tetra-specific antibody were the same.

GNC proteins are composed of Moiety 1 for binding at least one surfacemolecule on a T cell and Moiety 2 for binding at least one surfaceantigen on a cancer cell (TABLE 1A). The tetra-specific GNC antibodiescan be used to directly engage the body's endogenous T cells to killtumor cells independent of tumor antigen presentation by MHC to theantigen specific T cell receptors. This is in contrast to therapiesbased solely on immune checkpoint blockade, which have been limited byantigen recognition. In context, the immune checkpoint modulatingcomponent may be constructed as a part of tetra-specific GNC antibodies,which may provide benefits similar to that in a standard checkpointblockade therapy.

In addition to T cells, other cytotoxic cells may also be targeted byGNC proteins for cancer killing or preventing purposes. TABLE 1B showsthe example compositions of functional moieties (Moiety 1 and Moiety 2)and antigen binding domain in GNC proteins with NK cell binding domains.TABLE 1C shows the example compositions of functional moieties (Moiety 1and Moiety 2) and antigen binding domain in GNC proteins with macrophagebinding domains. TABLE 1D shows the example compositions of functionalmoieties (Moiety 1 and Moiety 2) and antigen binding domain in GNCproteins with dendritic cell binding domains.

GNC proteins are constructed to acquire multiple AgBDs specifically forbinding unequal numbers of T cell antagonists and agonists. In this way,GNC proteins may re-direct activated T cells to tumor cells with certainlevels of control of their activity in vivo (TABLE 2). Therefore, GNCproteins may be bi-specific, tri-specific, tetra-specific,penta-specific, hexa-specific, hepta-specific, or even octa-specificproteins. In the present invention, three classes of tetra-specific GNCantibodies, i.e. SI-39E, SI-35E, and SI-38E, were created to enableGNC-T cell therapy, of which antibody domains and its specificity islisted in TABLE 3. The structures of tetra-specific GNC antibodiestargeting EGFRvIII (SI-39E), ROR1 (SI-35E), and CD19 (SI-38E) are listedin TABLE 4.

Example 2: GNC-Activated, PBMS-Derived Cell Composition

The SI-35 class listed in Table 4 were tested for their ability toactivate and induce proliferation of different cell types, such as CD4+and/or CD8+ T cells and/or CD56+ natural killer cells (NK) within PBMC.The tetra-specific GNC antibodies were prepared at 2× finalconcentration and titrated in 1:10 serial dilutions across 6 wells of a96 well plate in 200 ul of RPMI+10% FBS. Human PBMC were purified bystandard Ficoll density gradient from a “leukopak” which is an enrichedleukapheresis product collected from normal human peripheral blood. Inthe final destination 96 well plate, the PBMC and serially titrated GNCproteins were combined by adding 100 μL of PBMC (100,000), and 100 μL ofeach antibody dilution to each well of the assay. The assay plate wasincubated at 37° C. for approximately 72 hours and then the contents ofeach assay well were harvested and analyzed by FACS for the number ofCD4+ T cells, CD8+ T cells, and CD56+NK cells. Cells were harvested fromeach well and transferred to a new 96 well V-bottom plate thencentrifuged at 400×g for 3 minutes. Supernatant was transferred to a 96well plate for analysis of IL-2 and Granzyme B. Cells were re-suspendedin 200 μL of 2% FBS/PBS of FACS antibodies and incubated on ice for 30minutes. The plate was centrifuged at 400×g for 3 minutes and thesupernatant was aspirated. This wash step was repeated once more andthen the cells were re-suspended in 100 μL 2% FBS/PBS and analyzed on aBD LSR FORTESSA.

As shown in FIG. 8, all SI-35E tetra-specific GNC antibodies, with theexception of those that had the scFv binding domain replaced with FITCat positions 2 (SI-35E37) and 4 (SI-35E39), induced production of IL-2from PBMC. These two proteins lacked the binding domains for PD-L1 orCD3 respectively. The secretion of Granzyme B into the culturesupernatant followed a similar pattern as that for IL-2 production asshown in FIG. 9. Both SI-35E37 and SI-35E3 were also much less potent atinducing cell-surface expression of the activation marker CD69 onCD4+(FIG. 10), CD8+(FIG. 11), and CD56+(FIG. 12) cells in the PBMCculture. Surface expression of the cytotoxic degranulation marker CD107a(LAMP-1) was induced by all GNC proteins tested except those lackingbinding at positions 2 and 4 on CD4+(FIG. 13), CD8+(FIG. 14), but lessconsistently on CD56+(FIG. 15) in the culture. At lower concentrations,3 of the GNC proteins (SI-35E42, SI-35E43, and SI-35E46) inducedexpression of CD69 on CD4+ T cells, CD8+ T cells, and CD56+NK cells,which correlated well with the level of IL-2 and granzyme B secretion(FIGS. 8 and 9) induced by these GNC.

Proliferation and production of gamma interferon was measured fromcultures of CD3+ or naïve CD8+ T cells (70,000 cells/well) stimulatedfor 5 days with a panel of SI-35 class antibodies. Human CD3+ orCD8+CD45RA+naïve T cells were enriched from peripheral blood mononuclearcells from a normal donor using the EasySep™ Human CD3+ or Naïve CD8+ TCell Isolation Kits (StemCell Technologies) as per the manufacturerprotocols. The final cell populations were determined to be >98% CD3+ orCD8+CD45RA+ T cells by flow cytometry. Proliferation in the culture wasmeasured after stain with Alamar blue (ThermoFisher Cat. No. DAL1100)for 1 hour at 37° C., and then read on a Spectramax plus 384 well reader(Molecular Devices). Proliferation of GNC-expanded CD3+ T cells wasexpressed as a fold increase in cell number over background of CD3+ Tcells in cell culture without GNC (FIG. 16). Proliferation was inducedby all constructs tested except the one lacking CD3 binding domain.Culture supernatants were also collected from these cultures andanalyzed for the presence of gamma interferon by ELISA. Secretion ofgamma interferon (FIG. 17) was high unless CD3 or ROR1 binding domainswere changed to FITC in the GNC constructs. Proliferation of naïveCD8+CD45RA+ T cells (FIG. 18) was more sensitive to the presence orabsence of 4-1BB binding domain compared to total CD3+ T cells as shownby addition of soluble anti-4-1BB monoclonal antibody to the culture inwhich 4-1BB binding on the GNC was absent. A similar pattern was foundfor secretion of gamma interferon from the naïve CD8+ T cells (FIG. 19).

Example 3. Scale Up and Formulation of a First GNC-Activated TherapeuticCell Composition

The manufacture of GNC-activated and -coated T cells at clinicallysignificant dosage of 10E9 was achieved after 7 days culture. Human PBMCwere isolated from LRS cone leukocytes by standard Ficoll densitygradient from leukopaks which are enriched leukapheresis productcollected from normal human peripheral blood. After collection the cellswere frozen at −80° C. and then later thawed before putting in culture.Using the G-Rex plate and bioreactor culture systems, the growth ofSI-38E17 GNC-stimulated PBMC cultures was monitored for up to 14 days.The culture medium consisted of RPMI 1640, 10% fetal calf serum, 1%non-essential amino acids, 1% GlutaMax, 0.6% glutamine-alaninesupplement, 15 ng/mL human IL-2, and 1 nM GNC protein. The 6-well G-Rexcultures tolerated seeding densities of 25-100 million PBMC/well for sixdays, which greatly exceeded recommended amounts, but was tolerated bythe cells in the system with a single 50% medium change on day 7.Clustering of cells was indicative of their activation in the culture(FIG. 20). At least 250 million cells from one leukapheresis donor wereseeded into two G-Rex 100M bioreactors and cultured in 1 liter ofculture medium for seven days. The larger volume of medium allowed theculture to continue without needing to exchange the culture medium. Cellyield in each of the 100M bioreactors was between 1.2-1.4 billion cellswith greater than 88% viability.

Example 4. A Second GNC-Activated Therapeutic Cell Composition

The cells from the bioreactor were harvested as the first GNC-activatedtherapeutic cell composition, which were optionally concentrated usingLOVO Automated Cell Processing System (Fresenius Kabi). One sample(Product B) was exposed to 1 nM SI-38E17, which is identical to thefirst GNC in this case for preparing a second GNC-activated therapeuticcell composition, potential for being used to target treat patientsharboring CD19 positive malignancies (FIG. 21A).

After the second concentration step (100 mL volume) during theprocessing in the LOVO system, the second GNC-activated therapeuticcells were washed twice before eluting to a final volume of 54 mL in asterile processing bag. The other sample (Product A) was only exposed tothe first GNC protein during the culture phase and not re-exposed duringprocessing in the LOVO system (FIG. 21A). Cells were removed from bags,mixed 1:1 with CryoStor CS10 reagent, and frozen to −80° C. Theprocessed cells were thawed and compared to the thawed unstimulated PBMCfrom the same donor before culture.

Cell viability from the GNC-expanded T cell (GET) culture was >75% andwas not affected by exposure to additional GNC reagent (GNC-T, ProductB) during processing (FIG. 21B). The mean diameter of the cellsincreased during culture, indicative of cell activation. Flow cytometrywas performed on the input PBMC cell material and the two formulationsafter thawing using a multi-color panel of antibodies to stain for:live/dead (e780), CD45, TCRα/β, CD56, CD4, CD8, CD14, TCRγ/δ, and CD20.Gating for quantification of the different cell subsets is shown on theGNC-activated T cells (Product A) and the additional GNC-coated GNC-Tcells (Product B) (FIGS. 22A and 22B). The percentages of eachsubpopulation of cells were similar between Product A and Product B, butvery different from those of input PBMC (FIG. 22C). FIG. 23 summariesthe total number and percentage of each subpopulation of cells. Comparedto the input PBMC cell material, while the total number of leukocytesincreased from 250 to 1000 millions or four-fold, the total number ofeach subpopulation of T cells was vastly increased by 55-fold for α/β Tcells, 45-fold for CD4+ T cells, and 78-fold for CD8+ T cells. In thiscontext, the increase of γ/δ T cells was modest at 5-fold, andTCRα/β−/lo, γ/δ+, CD8+ T cells seemed to the most abundant. Finally, thecharacteristic feature of both Product A and Product B cell compositionsis the fact that there were no detectable B cells.

This example illustrates a number of advantages of GNC-T cells incomparison to CAR-T cell preparations. First, the cell composition ofthe starting material was fresh PBMC from the donor and did not need tobe pre-selected for particular subsets of cells or require addition offeeder cells or synthetic beads. The GNC protein was 100% non-nucleotidebiological material, and did not require the transfer of RNA or DNA intothe cells, or transfection with a viral vector. The GNC-inducedexpansion yielded a therapeutic dose in 9 days, compared to the averageof 40 days for CAR-T cell expansion. The resulting cells were devoid ofB cells and highly enriched for activated CD4+ and CD8+ T cells that hadpotent killing potential against their specific targets. The GNCtherapeutic composition was viable and bioactive upon thaw from −80° C.Together these advantages are expected to significantly lower waitingtimes, costs and issues related to infrastructure and training relatedto CAR-T cell therapy. Improvements in the purity, safety and quantityof the end product will be of significant benefit to the patient.

Example 5. PBMC Pre-Activated with GNC Proteins are Redirected toPotently Kill Tumor Cells

Six of the GNC SI-35 class proteins listed in Table 4 were tested forthe ability to activate PBMC for redirected T cell cytotoxicity (RTCC)activity against a human ROR1-transduced CHO cell line (FIG. 24). GNCproteins were prepared at 2× final concentration and titrated 1:3 across10 wells of a 96 well plate in 200 ul of RPMI+10% FBS. In the finaldestination 96 well plate, the PBMC and serially titrated antibodieswere combined by adding 100 μL of PBMC (200,000), and 100 μL of eachantibody dilution to each well of the assay. The assay plate wasincubated at 37° C. for approximately 72 hours before the addition ofCFSE-labeled CHO-ROR1 cells. CHO-ROR1 target cells, 5×10e6, were labeledwith CFSE (Invitrogen, #C34554) at 0.5 μM in 10 mL of culture media for20 minutes at 37° C. The CHO-ROR1 cells were washed 3 times with 50 mLof culture media before resuspending in 10 mL, counted again and then5,000 CFSE-labeled CHO-ROR1 cells were added to each well ofGNC-activated PBMC. Cells were incubated for another 72 hours and thenthe contents of each assay well were harvested and analyzed for thenumber of CFSE-labeled target cells remaining. As shown on FIG. 24, allof the GNC proteins tested directed RTCC activity with SI-35E42,SI-35E43, and SI-35E46 being the most potent in reducing the number ofCHO-ROR1 cells in the well.

To further demonstrate the killing effects of GNC-labeled PBMC againsthuman tumor cells, a GNC-dose and effector:target ratio escalationexperiment was performed using an IncuCyte S3 Live Cell Analysis System(Sartorius/Essen Biosciences) to monitor the cells over time. PBMC froma healthy donor were labeled with GNC protein SI-38E17 at 10-fold serialdoses ranging from 0.01 to 100 nM for 30 minutes at 37° C. and thenwashed prior to culture. The GNC SI-38E17 targets the CD19 antigenexpressed on B cell surfaces, and therefore, the Kasumi-2 precursor Bcell leukemia line was chosen as a target cell. The Kasumi-2 cell usedwas transduced to express green fluorescence protein (GFP) and thereforethe presence of tumor cells was tracked by measuring the average greenfluorescence in 4 images/well collected 9 times over a six-day period.The effector:target (E:T) ratios were escalated by adding GNC-labeledPBMC in a serial 2-fold dilution of 5,000 (1:1) to 160,000 (32:1) cellsto duplicate wells. As shown in FIG. 25, Kasumi-2 cells increased innumber in the wells that had from 1:1 to 8:1 E:T ratios of unlabeledPBMC. Exposure to as little as 0.1 nM GNC led to decreased growth ofKasumi-2 in the 1:1 culture with suppression increasing at each 2-foldincrease in the E:T ratio. Coating of PBMC with 1 nM or greaterconcentrations of GNC led to nearly complete elimination of Kasumi-2cells after 42 hours of culture at all E:T ratios.

As a follow up experiment, three other transformed B cell lines: NALM-6,MEC-1, and Daudi and the acute T cell leukemia line, Jurkat, were usedas target cells. These target cells were previously transduced bylentivirus to constitutively express the NucRed 647 molecule. In thisassay, PBMC were exposed to 10-fold doses of GNC protein SI-38E17 for 30minutes at 37° C. and then washed as before. PBMC were plated at 1.2×10⁶cells/well and 50,000 target tumor cells were added. Cells were placedin IncuCyte S3 set to collect red fluorescence images (4 images/well)collected at 10 time points over a 5.5-day period (FIG. 26). Growthcurves were established for all four tumor cell lines in the absence ofPBMC (null). Labeling of PBMC with 1 nM or more of GNC protein SI-38E17led to arrested growth of all three B cell lines but not Jurkat T cellleukemia. The B cell lines varied in their susceptibility to PBMC cellspre-exposed to 0.1 nM of GNC protein.

As a different method of quantifying the outcome of cultures of GNC-Tcells with tumor cells, we established a limit of quantification (LOQ)curve for detection by flow cytometry. Daudi-Red cells were seriallydiluted 10-fold in a range from 200,000 to 20 cells and then mixed 1:1with 1 million PBMC to create samples of 10%, 1.0%, 0.1%, 0.01% and0.001% tumor cells, which were then analyzed by flow cytometry (FIG.27). Next, cells were harvested from a 15 day 6-well G-Rex culture of 1nM GNC-expanded T cells that had been spiked with 10%, 1% or 0.1% ofNALM-6, MEC-1, Daudi, or Jurkat (all NucRed-transduced) tumor cells attime 0 and analyzed using the same flow cytometry settings as above.Tumor cells were reduced to less than 0.001% in all conditions with theexception of the culture in which the MEC-1 tumor line was spiked in at10% were 44 cells were detected. In this condition the MEC-1 cells werereduced to <0.01% in the culture.

While the present disclosure has been described with reference toparticular embodiments or examples, it may be understood that theembodiments are illustrative and that the disclosure scope is not solimited. Alternative embodiments of the present disclosure may becomeapparent to those having ordinary skill in the art to which the presentdisclosure pertains. Such alternate embodiments are considered to beencompassed within the scope of the present disclosure. Accordingly, thescope of the present disclosure is defined by the appended claims and issupported by the foregoing description. All references cited or referredto in this disclosure are hereby incorporated by reference in theirentireties.

Tables

TABLE 1A Composition of example GNC proteins with T cell bindingdomains. Moiety 1 Activation Agonist Moiety 2 of T cells receptorAntagonist receptor Tumor Antigen CD3 CD28, 41BB, PDL1, PD1, TIGIT,BCMA, CD19, CD20, OX40, GITR, TIM-3, LAG-3, CD33, CD123, CD22, CD40L,ICOS, CTLA4, BTLA, CD30, ROR1, CEA, Light, CD27, VISTA, PDL2 HER2, EGFR,CD30 EGFRvIII, LMP1, LMP2A, Mesothelin, PSMA, EpCAM, glypican-3, gpA33,GD2, TROP2

TABLE 2 Examples of possible combinations of T cell activation, T cellagonist, T cell antagonist, and tumor antigen binding domains in asingle GNC protein. T cell Tumor T cell T cell T cell T cell T cell Tcell GNC protein activation antigen antagonist agonist antagonistantagonist antagonist agonist Bi-specific CD3 ROR1 Tri-specific CD3 ROR1PD1 Tetra-specific CD3 ROR1 PD1 41BB Penta-specific CD3 ROR1 PD1 41BBLAG3 Hexa-specific CD3 ROR1 PD1 41BB LAG3 TIM3 Hepta-specific CD3 ROR1PD1 41BB LAG3 TIM3 TIGIT Octa-specific CD3 ROR1 PD1 41BB LAG3 TIM3 TIGITCD28

TABLE 3 Specificity of antibody binding domains used in GNC proteins.AgBD Specificity Antibody Name CD3ε 284A10 480C8 4-1BB 460C3 420H5 466F6FITC 4420 PD-L1 PL230C6 CD19 21D4 ROR1 323H7 IgD Domain 330F11 KringleDomain 338H4 Frizzled Domain 324C6 EGFRvIII 806

TABLE 4 Classes of tetra-specific GNC antibodies targeting EGFRvIII(SI-39E), ROR1 (SI-35E), and CD19 (SI-38E). GNC AgBD 1 Humanized AgBD 2Humanized IgG1 AgBD 3 Humanized AgBD 4 Humanized ID (LH-scFv) Variant(Fab) Variant Fc (HL-scFv) Variant (HL-ScFv) Variant SI-39E18 284A10L1H1 806 — n2 PL221G5 H1L1 420H5 H3L3 SI-39E29 806 — 284A10 H1L1 n2PL221G5 H1L1 420H5 H3L3 SI-35E20 466F6 L5H2 PL230C6 H3L2 n2 323H7 H4L1284A10 H1L1 SI-35E58 284A10 L1H1 PL230C6 H3L2 n2 323H7 H4L1 466F6 H2L5SI-35E88 284A10 L1H1 323H7 H4L1 n2 PL230C6 H3L2 466F6 H2L5 SI-35E99284A10 L1H1 323H7 H4L1 n2 PL221G5 H1L1 466F6 H2L5 SI-35E18 460C3 L1H1PL230C6 H3L2 n2 323H7 H4L1 284A10 H1L1 SI-35E19 420H5 L3H3 PL230C6 H3L2n2 323H7 H4L1 284A10 H1L1 SI-35E36 4420 — PL230C6 H3L2 n2 338H4 H3L4284A10 H1L1 SI-35E37 460C3 L1H1 4420 — n2 338H4 H3L4 284A10 H1L1SI-35E38 460C3 L1H1 PL230C6 H3L2 n2 4420 — 284A10 H1L1 SI-35E39 460C3L1H1 PL230C6 H3L2 n2 338H4 H3L4 4420 — SI-38E17 284A10 H1L1 21D4 — n2PL221G5 H1L1 466F6 H2L5 SI-38E33 21D4 — 284A10 H1L1 n2 PL221G5 H1L1466F6 H2L5

SEQ ID Description 1 anti-CD3 284A10 VHv1 nt 2 anti-CD3 284A10 VHv1 aa 3anti-CD3 284A10 VLv1 nt 4 anti-CD3 284A10 VLv1 aa 5 anti-PD-L1 PL23006VHv3 nt 6 anti-PD-L1 PL23006 VHv3 aa 7 anti-PD-L1 PL23006 VLv2 nt 8anti-PD-L1 PL23006 VLv2 aa 9 anti-PD-L1 PL221G5 VHv1 nt 10 anti-PD-L1PL221G5 VHv1 aa 11 anti-PD-L1 PL221G5 VLv1 nt 12 anti-PD-L1 PL221G5 VLv1aa 13 anti-4-1BB 420H5 VHv3 nt 14 anti-4-1BB 420H5 VHv3 aa 15 anti-4-1BB420H5 VLv3 nt 16 anti-4-1BB 420H5 VHLv3 aa 17 anti-4-1BB 466F6 VHv2 nt18 anti-4-1BB 466F6 VHv2 aa 19 anti-4-1BB 466F6 VLv5 nt 20 anti-4-1BB466F6 VLv5 aa 21 anti-4-1BB 460C3 VHv1 nt 22 anti-4-1BB 460C3 VHv1 aa 23anti-4-1BB 460C3 VLv1 nt 24 anti-4-1BB 460C3 VLv1 aa 25 anti-ROR1 323H7VHv4 nt 26 anti-ROR1 323H7 VHv4 aa 27 anti-ROR1 323H7 VLv1 nt 28anti-ROR1 323H7 VLv1 aa 29 anti-ROR1 338H4 VHv3 nt 30 anti-ROR1 338H4VHv3 aa 31 anti-ROR1 338H4 VLv4 nt 32 anti-ROR1 338H4 VLv4 aa 33anti-FITC 4-4-20 VH nt 34 anti-FITC 4-4-20 VH aa 35 anti-FITC 4-4-20 VLnt 36 anti-FITC 4-4-20 VL aa 37 human IgG1 null2 (G1m-fa with ADCC/CDCnull mutations) nt 38 human IgG1 null2 (G1m-fa with ADCC/CDC nullmutations) aa 39 human Ig Kappa nt 40 human Ig Kappa aa 41 SI-35E18(460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv × 284A10-H1L1-scFv)heavy chain nt 42 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab ×323H7-H4L1-scFv × 284A10-H1L1-scFv) heavy chain aa 43 SI-35E18(460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv × 284A10-H1L1-scFv)light chain nt 44 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab ×323H7-H4L1-scFv × 284A10-H1L1-scFv) light chain aa 45 anti-CD3 284A10VHv1b nt 46 anti-CD3 284A10 VHv1b aa 47 anti-huCD19 21D4 VH nt 48anti-huCD19 21D4 VH aa 49 anti-huCD19 21D4 VL nt 50 anti-huCD19 21D4 VLaa 51 anti-huEGFRvIII 806 VH nt 52 anti-huEGFRvIII 806 VH aa 53anti-huEGFRvIII 806 VL nt 54 anti-huEGFRvIII 806 VL aa 55 GGGGSGGGGSGlinker nt 56 GGGGSGGGGSG linker aa 57 GGGSGGGGS linker 01 nt 58GGGSGGGGS linker 01 aa 59 GGGSGGGGS linker 02 nt 60 GGGSGGGGS linker 02aa 61 GGGSGGGGSGGGSGGGGS linker nt 62 GGGSGGGGSGGGSGGGGS linker aa 63SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-sc Fv ×420H5-H3L3-scFv) heavy chain nt 64 SI-39E18 (284A10-L1H1-scFv × 806-Fab× PL221G5-H1L1-sc Fv × 420H5-H3L3-scFv) heavy chain aa 65 SI-39E18(284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-sc Fv × 420H5-H3L3-scFv)light chain nt 66 SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain aa 67 SI-39E29 (806-LH-scFv ×284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) heavy chain nt 68SI-39E29 (806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv ×420H5-H3L3-scFv) heavy chain aa 69 SI-39E29 (806-LH-scFv × 284A10-Fab ×PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain nt 70 SI-39E29(806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) lightchain aa 71 SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 3 23H7-H4L1-scFv ×284A10-H1L1-scFv) heavy chain nt 72 SI-35E20 (466F6-L5H2-scFv ×PL230C6-Fab × 323H7-H4L1-scFv × 284A10-H1L1-scFv) heavy chain aa 73SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) light chain nt 74 SI-35E20 (466F6-L5H2-scFv ×PL230C6-Fab × 323H7-H4L1-scFv × 284A10-H1L1-scFv) light chain aa 75SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-sc Fv ×466F6-H2L5-scFv) heavy chain nt 76 SI-35E58 (284A10-L1H1-scFv ×PL230C6-Fab × 323H7-H4L1-sc Fv × 466F6-H2L5-scFv) heavy chain aa 77SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-sc Fv ×466F6-H2L5-scFv) light chain nt 78 SI-35E58 (284A10-L1H1-scFv ×PL230C6-Fab × 323H7-H4L1-sc Fv × 466F6-H2L5-scFv) light chain aa 79SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-sc Fv ×466F6-H2L5-scFv) heavy chain nt 80 SI-35E88 (284A10-L1H1-scFv ×323H7-Fab × PL230C6-H3L2-sc Fv × 466F6-H2L5-scFv) heavy chain aa 81SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-sc Fv ×466F6-H2L5-scFv) light chain nt 82 SI-35E88 (284A10-L1H1-scFv ×323H7-Fab × PL230C6-H3L2-sc Fv × 466F6-H2L5-scFv) light chain aa 83SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-sc Fv ×466F6-H2L5-scFv) heavy chain nt 84 SI-35E99 (284A10-L1H1-scFv ×323H7-Fab × PL221G5-H1L1-sc Fv × 466F6-H2L5-scFv) heavy chain aa 85SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-sc Fv ×466F6-H2L5-scFv) light chain nt 86 SI-35E99 (284A10-L1H1-scFv ×323H7-Fab × PL221G5-H1L1-sc Fv × 466F6-H2L5-scFv) light chain aa 87SI-38E17 (284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv ×466F6-H2L5-scFv) heavy chain nt 88 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab× PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain aa 89 SI-38E17(284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv)light chain nt 90 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab ×PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain aa 91 SI-38E33(21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-sc Fv × 466F6-H2L5-scFv) heavychain nt 92 SI-38E33 (21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-sc Fv ×466F6-H2L5-scFv) heavy chain aa 93 SI-38E33 (21D4-LH-scFv × 284A10-Fab ×PL221G5-H1L1-sc Fv × 466F6-H2L5-scFv) light chain nt 94 SI-38E33(21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-sc Fv × 466F6-H2L5-scFv) lightchain aa

GNC-T Sequence Listing of Tetra-Specific GNC Antibodies

>SEQ ID 01 anti-CD3 284A10 VHv1 ntGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCA >SEQ ID 02 anti-CD3 284A10 VHv1 aaEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSS >SEQ ID 03 anti-CD3 284A10 VLv1 ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 04 anti-CD3 284A10 VLv1 aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIK >SEQ ID 05 anti-PD-L1 PL230C6 VHv3 ntCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT >SEQ ID 06 anti-PD-L1 PL230C6 VHv3 aaQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGIITYSGSRYYANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSS >SEQ ID 07 anti-PD-L1 PL230C6 VLv2 ntGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 08 anti-PD-L1 PL230C6 VLv2 aaAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIK >SEQ ID 09 anti-PD-L1 PL221G5 VHv1 ntGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC >SEQ ID 10 anti-PD-L1 PL221G5 VHv1 aaEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSS >SEQ ID 11 anti-PD-L1 PL221G5 VLv1 ntGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 12 anti-PD-L1 PL221G5 VLv1 aaDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIK >SEQ ID 13 anti-4-1BB 420H5 VHv3 ntCAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC >SEQ ID 14 anti-4-1BB 420H5 VHv3 aaQSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWICWVRQAPGKGLEWIACIYVGSSGDTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNLWGQGTLVTVSS >SEQ ID 15 anti-4-1BB 420H5 VLv3 ntGCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTATGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 16 anti-4-1BB 420H5 VLv3 aaALVMTQSPSTLSASVGDRVTINCQASEDIDTYLAWYQQKPGKAPKLLIFYASDLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGAFGGGTKVEIK >SEQ ID 17 anti-4-1BB 466F6 VHv2 ntCGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC >SEQ ID 18 anti-4-1BB 466F6 VHv2 aaRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSS >SEQ ID 19 anti-4-1BB 466F6 VLv5 ntGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 20 anti-4-1BB 466F6 VLv5 aaDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK >SEQ ID 21 anti-4-1BB 460C3 VHv1 ntGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGAATCGACTTCAGTAGGAGATACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATATATACTGGTAGCCGCGATACTCCTCACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGAAGGTAGCCTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC >SEQ ID 22 anti-4-1BB 460C3 VHv1 aaEVQLLESGGGLVQPGGSLRLSCAASGIDFSRRYYMCWVRQAPGKGLEWIACIYTGSRDTPHYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREGSLWGQGTLVTVSS >SEQ ID 23 anti-4-1BB 460C3 VLv1 ntGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAGTAACTGGTTCTCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCGCAGGCGGTTACAATACTGTTATTGATACTTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 24 anti-4-1BB 460C3 VLv1 aaDIQMTQSPSTLSASVGDRVTITCQSSQSVYSNWFSWYQQKPGKAPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGYNTVIDTFAFGGGTKVEIK >SEQ ID 25 anti-ROR1 323H7 VHv4 ntGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA >SEQ ID 26 anti-ROR1 323H7 VHv4 aaEVQLLESGGGLVQPGGSLRLSCAASGETISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSS >SEQ ID 27 anti-ROR1 323H7 VLv1 ntGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 28 anti-ROR1 323H7 VLv1 aaDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIK >SEQ ID 29 anti-ROR1 338H4 VHv3 ntGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAGGGGGCTGGAGTGGATCGGAATCATTTATGCTAGTGGTAGCACATACTACGCGAGCTCGGCGAAAGGCAGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAATTTATGACGGCATGGACCTCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA >SEQ ID 30 anti-ROR1 338H4 VHv3 aaEVQLVESGGGLVQPGGSLRLSCTASGFSLSSYAMSWVRQAPGRGLEWIGIIYASGSTYYASSAKGRFTISKDNTKNTVDLQMNSLRAEDTAVYYCARIYDGMDLWGQGTLVTVSS >SEQ ID 31 anti-ROR1 338H4 VLv4 ntGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAACATTTACAGCTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCGCCTGATCTATCTGGCATCTACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTACACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAGCAATTATAACGGTAATTATGGTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 32 anti-ROR1 338H4 VLv4 aaDIQMTQSPSSLSASVGDRVTINCQASQNIYSYLSWYQQKPGKVPKRLIYLASTLASGVPSRFSGSGSGTDYTLTISSLQPEDVATYYCQSNYNGNYGFGGGTKVEIK >SEQ ID 33 anti-FITC 4420 VH ntGAGGTGAAGCTGGATGAGACTGGAGGAGGCTTGGTGCAACCTGGGAGGCCCATGAAACTCTCCTGTGTTGCCTCTGGATTCACTTTTAGTGACTACTGGATGAACTGGGTCCGCCAGTCTCCAGAGAAAGGACTGGAGTGGGTAGCACAAATTAGAAACAAACCTTATAATTATGAAACATATTATTCAGATTCTGTGAAAGGCAGATTCACCATCTCAAGAGATGATTCCAAAAGTAGTGTCTACCTGCAAATGAACAACTTAAGAGTTGAAGACATGGGTATCTATTACTGTACGGGTTCTTACTATGGTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA >SEQ ID 34 anti-FITC 4420 VH aaEVKLDETGGGLVQPGRPMKLSCVASGFTFSDYWMNWVRQSPEKGLEWVAQIRNKPYNYETYYSDSVKGRFTISRDDSKSSVYLQMNNLRVEDMGIYYCTGSYYGMDYWGQGTSVTVSS >SEQ ID 35 anti-FITC 4420 VL ntGATGTCGTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCCTTGTACACAGTAATGGAAACACCTATTTACGTTGGTACCTGCAGAAGCCAGGCCAGTCTCCAAAGGTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGAGTGGAGGCTGAGGATCTGGGAGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA >SEQ ID 36 anti-FITC 4420 VL aaDVVMTQTPLSLPVSLGDQASISCRSSQSLVHSNGNTYLRWYLQKPGQSPKVLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPWTFGGGTKLEIK >SEQ ID 37 human IgG1 null (G1m-fa with ADCC/CDC null mutations) ntGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT >SEQ ID 38 human IgG1 null (G1m-fa with ADCC/CDC null mutations) aaASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG >SEQ ID 39 human Ig Kappa ntCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 40 human Ig Kappa aaRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 41 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) heavy chain ntGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAGTAACTGGTTCTCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCGCAGGCGGTTACAATACTGTTATTGATACTTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGAATCGACTTCAGTAGGAGATACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATATATACTGGTAGCCGCGATACTCCTCACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGAAGGTAGCCTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 42 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) heavy chain aaDIQMTQSPSTLSASVGDRVTITCQSSQSVYSNWFSWYQQKPGKAPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGYNTVIDTFAFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGIDFSRRYYMCWVRQAPGKGLEWIACIYTGSRDTPHYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREGSLWGQGTLVTVSSGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGIITYSGSRYYANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGETISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIK >SEQ ID 43 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) light chain ntGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 44 SI-35E18 (460C3-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) light chain aaAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 45 anti-CD3 284A10 VHv1b ntGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACA >SEQ ID 46 anti-CD3 284A10 VHv1b aaEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVST >SEQ ID 47 anti-huCD19 21D4 VH ntGAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA >SEQ ID 48 anti-huCD19 21D4 VH aaEVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIGWVRQAPGKGLEWMGIIYPDDSDTRYSPSFQGQVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDFWGQGTLVTVSS >SEQ ID 49 anti-huCD19 21D4 VL ntGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAA >SEQ ID 50 anti-huCD19 21D4 VL aaAIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFTFGPGTKVDIK >SEQ ID 51 anti-huEGFRvIII 806 VH ntGATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA >SEQ ID 52 anti-huEGFRvIII 806 VH aaDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSA >SEQ ID 53 anti-huEGFRvIII 806 VL ntGACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA >SEQ ID 54 anti-huEGFRvIII 806 VL aaDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK >SEQ ID 55 GGGGSGGGGSG linker ntGGCGGTGGAGGGTCCGGCGGTGGTGGCTCCGGA >SEQ ID 56 GGGGSGGGGSG linker aaGGGGSGGGGSG >SEQ ID 57 GGGGSGGGGS linker 01 ntGGCGGTGGAGGGTCCGGCGGTGGTGGATCA >SEQ ID 58 GGGGSGGGGS linker 01 aaGGGGSGGGGS >SEQ ID 59 GGGGSGGGGS linker 02 ntGGCGGTGGAGGGTCCGGCGGTGGTGGATCC >SEQ ID 60 GGGGSGGGGS linker 02 aaGGGGSGGGGS >SEQ ID 61 GGGGSGGGGSGGGGSGGGGS linker ntGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCA >SEQ ID 62 GGGGSGGGGSGGGGSGGGGS linker aaGGGGSGGGGSGGGGSGGGGS >SEQ ID 63 SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) heavy chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGTATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTACGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 64 SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) heavy chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIKGGGGSGGGGSQSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWICWVRQAPGKGLEWIACIYVGSSGDTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSALVMTQSPSTLSASVGDRVTINCQASEDIDTYLAWYQQKPGKAPKLLIFYASDLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGAFGGGTKVEIK >SEQ ID 65 SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain ntGACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 66 SI-39E18 (284A10-L1H1-scFv × 806-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain aaDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 67 SI-39E29 (806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) heavy chain ntGACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGTATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTACGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 68 SI-39E29 (806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) heavy chain aaDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFIISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIKGGGGSGGGGSQSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWICWVRQAPGKGLEWIACIYVGSSGDTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSALVMTQSPSTLSASVGDRVTINCQASEDIDTYLAWYQQKPGKAPKLLIFYASDLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGAFGGGTKVEIK >SEQ ID 69 SI-39E29 (806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 70 SI-39E29 (806-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 420H5-H3L3-scFv) light chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 71 SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) heavy chain ntGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 72 SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) heavy chain aaDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGIITYSGSRYYANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIK >SEQ ID 73 SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) light chain ntGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 74 SI-35E20 (466F6-L5H2-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×284A10-H1L1-scFv) light chain aaAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 75 SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×466F6-H2L5-scFv) heavy chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACCGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCArCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACrCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGArCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTrATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 76 SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×466F6-H2L5-scFv) heavy chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSTGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGsIITYSGSRYY+EE ANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK >SEQ ID 77 SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×466F6-H2L5-scFv) light chain ntGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 78 SI-35E58 (284A10-L1H1-scFv × PL230C6-Fab × 323H7-H4L1-scFv ×466F6-H2L5-scFv) light chain aaAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 79 SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-scFv × 466F6-H2L5-scFv) heavy chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACCGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCCGGTGGAGGCGGTTCAGGCGGAGGTGGAAGTGGTGGTGGCGGCTCTGGAGGCGGCGGATCTGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 80 SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-scFv × 466F6-H2L5-scFv) heavy chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSTGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGIITYSGSRYYANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK >SEQ ID 81 SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-scFv × 466F6-H2L5-scFv) light chain ntGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 82 SI-35E88 (284A10-L1H1-scFv × 323H7-Fab × PL230C6-H3L2-scFv × 466F6-H2L5-scFv) light chain aaDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVIEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 83 SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGCTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 84 SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSTGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK >SEQ ID 85 SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain ntGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 86 SI-35E99 (284A10-L1H1-scFv × 323H7-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain aaDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVIEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 87 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 88 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSGGGGSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIGWVRQAPGKGLEWMGIIYPDDSDTRYSPSFQGQVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIKGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK >SEQ ID 89 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain ntGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT >SEQ ID 90 SI-38E17 (284A10-L1H1-scFv × 21D4-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain aaAIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >SEQ ID 91 SI-38E33 (21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain ntGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA >SEQ ID 92 SI-38E33 (21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) heavy chain aaAIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFTFGPGTKVDIKGGGGSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIGWVRQAPGKGLEWMGIIYPDDSDTRYSPSFQGQVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDFWGQGTLVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMCWVRQAPGKGLEWIACIAAGSAGITYDANWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDLWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLNWYQQKPGKAPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNVFGGGTKVEIKGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQWVRQAPGKGLEYIGTISSGGNVYYASSARGRFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPMWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLSWYQQKPGKAPKLLIYAAANLASGVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGAFGGGTKVEIK SEQ ID 93 SI-38E33 (21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain ntGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTSEQ ID 94 SI-38E33 (21D4-LH-scFv × 284A10-Fab × PL221G5-H1L1-scFv × 466F6-H2L5-scFv) light chain aaDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

What we claim is:
 1. A method for generating a therapeutic composition,comprising providing a cell material comprising a cytotoxic cell,incubating the cell material with a first GNC protein to provide anactivated cell composition, wherein the activated cell compositioncomprises a first therapeutic cell, wherein the first GNC proteincomprising a first cytotoxic binding moiety and a first cancer targetingmoiety, wherein the first cytotoxic binding moiety has a specificity toa first cytotoxic cell receptor and is configured to activate the firstcytotoxic cell through the binding with the first cytotoxic cellreceptor, and wherein the first cancer targeting moiety has aspecificity to a first cancer cell receptor, and wherein the firsttherapeutic cell comprises the first GNC protein bound to the cytotoxiccell through the binding interaction with the first cytotoxic cellreceptor, and formulating the activated cell composition to provide atherapeutic composition, wherein the therapeutic composition issubstantially free of exogenous viral and non-viral DNA or RNA.
 2. Themethod of claim 1, wherein the incubating step is repeated by incubatinga second GNC protein with the activated cell composition, wherein thesecond GNC protein comprising a second cytotoxic binding moiety and asecond cancer targeting moiety, wherein the second cytotoxic bindingmoiety has a specificity to a second cytotoxic cell receptor, andwherein the second cancer targeting moiety has a specificity to a secondcancer cell receptor, wherein the activated cell composition furthercomprises a second therapeutic cell, and wherein the second therapeuticcell comprises the second GNC protein bound to the cytotoxic cell or thefirst therapeutic cell through the binding interaction with the secondcytotoxic cell receptor.
 3. The method of claim 2, wherein the secondGNC protein is the same as the first GNC protein.
 4. The method of claim2, wherein the second GNC protein is different from the first GNCprotein.
 5. The method of claim 1, wherein the first or the secondcancer targeting moiety has the specificity against B cell, and whereinthe therapeutic composition is substantially free of B cell.
 6. Themethod of claim 1, wherein the cytotoxic cell receptor comprises aT-cell receptor, a NK cell receptor, a macrophage receptor, a dendriticcell receptor, or a combination thereof.
 7. The method of claim 1,wherein the molar to cell ratio between the first GNC protein and thecytotoxic cell is at least 30 to 1 when incubating the cell materialwith the first GNC protein.
 8. The method of claim 1, wherein thetherapeutic composition comprises at least 10⁶ cells per ml.
 9. Themethod of claim 1, wherein the therapeutic composition comprises thefirst therapeutic cell, the first GNC protein, the cytotoxic cell, or acombination thereof.
 10. The method of claim 2, wherein the therapeuticcomposition comprises the second therapeutic cell, the second GNCprotein, comprises the first therapeutic cell, the first GNC protein,the cytotoxic cell, or a combination thereof.
 11. The method of claim 1,wherein the cell material comprises PBMC.
 12. The method of claim 1,wherein the first and the second cancer-targeting moiety independentlyhas a specificity for CD19, PDL1, or a combination thereof.
 13. Themethod of claim 1, wherein the first and the second cytotoxic bindingmoiety independently has a specificity for CD3, PDL1, 41BB, or acombination thereof.
 14. A method of treating a subject having a cancer,comprising providing a cell material comprising a cytotoxic cell,incubating the cell material with a first GNC protein to provide anactivated cell composition, wherein the activated cell compositioncomprises a first therapeutic cell, wherein the first GNC proteincomprising a first cytotoxic binding moiety and a first cancer targetingmoiety, wherein the first cytotoxic binding moiety has a specificity toa first cytotoxic cell receptor and is configured to activate the firstcytotoxic cell through the binding with the first cytotoxic cellreceptor, and wherein the first cancer targeting moiety has aspecificity to a first cancer cell receptor, and wherein the firsttherapeutic cell comprises the first GNC protein bound to the cytotoxiccell through the binding interaction with the first cytotoxic cellreceptor, and formulating the activated cell composition to provide atherapeutic composition, wherein the therapeutic composition issubstantially free of exogenous viral and non-viral DNA or RNA, andadministering the therapeutic composition to the subject.
 15. The methodof claim 14, wherein the incubating step is repeated by incubating asecond GNC protein with the activated cell composition, wherein thesecond GNC protein comprising a second cytotoxic binding moiety and asecond cancer targeting moiety, wherein the second cytotoxic bindingmoiety has a specificity to a second cytotoxic cell receptor, andwherein the second cancer targeting moiety has a specificity to a secondcancer cell receptor, wherein the activated cell composition furthercomprises a second therapeutic cell, and wherein the second therapeuticcell comprises the second GNC protein bound to the cytotoxic cell or thefirst therapeutic cell through the binding interaction with the secondcytotoxic cell receptor.
 16. The method of claim 14, wherein the secondGNC protein is the same as the first GNC protein.
 17. The method ofclaim 14, wherein the second GNC protein is different from the first GNCprotein.
 18. The method of claim 14, wherein the first or the secondcancer targeting moiety has the specificity against B cell, and whereinthe therapeutic composition is substantially free of B cell.
 19. Themethod of claim 14, further comprising isolating the cytotoxic cell fromperipheral blood mononuclear cells (PBMC) before providing the cellmaterial.
 20. The method of claim 19, further comprising isolating theperipheral blood mononuclear cells (PBMC) from a blood. 21-22.(canceled)
 23. The method of claim 14, further comprising administeringan additional GNC protein to the subject after the administering thetherapeutic composition to the subject.
 24. The method of claim 14,wherein the cytotoxic cell comprises T cell, NK cell, or a combinationthereof.
 25. The method of claim 19, wherein the isolating the cytotoxiccell comprising isolating at least one subpopulation of cytotoxic cellto provide therapeutic T cells, wherein the subpopulation of cytotoxiccell comprises CD3+ cells, CD4+ cells, CD8+ cells, CD56+ cells, CD28+cells, CD69+ cells, CD107a+ cells, CD45RA+ cells, CD45RO+ cells, γδ TCR+cells, αβ TCR+ cells, CD25+ cells, CD127^(lo/−) cells, CCR7+ cells,PD-1+ cells or a combination thereof.
 26. (canceled)
 27. The method ofclaim 14, further comprising evaluating therapeutic efficacy after theadministering step, wherein the evaluating therapeutic efficacycomprises checking one or more biomarkers of the cancer, monitoring thelife span of the therapeutic cells, or a combination thereof. 28-29.(canceled)
 30. The method of claim 14, wherein the subject is a human.31. The method of claim 14, wherein the cancer comprises cellsexpressing ROR1, CEA, HER2, EGFR, EGFR VIII, LMP1, LMP2A, Mesothelin,PSMA, EpCAM, glypican-3, gpA33, GD2, TROP2, BCMA, CD19, CD20, CD33,CD123, CD22, CD30, or a combination thereof.
 32. (canceled)
 33. Themethod of claim 14, wherein the cancer is CD19 positive.
 34. The methodof claim 14, further comprising administering an effective amount of atherapeutic agent after the administering the therapeutic composition tothe subject.
 35. The method of claim 34, wherein the therapeutic agentcomprises a monoclonal antibody, a multi-specific antibody, achemotherapy agent, an enzyme, a protein, a co-stimulator, an apoptosissensitizer, a tumor vascular disruptor, or a combination thereof,wherein the co-stimulator is configured to increase the amount ofcytotoxic T cells in the subject. 36-39. (canceled)